Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-Vimentin RabMAb (ab92547, left panel) (brown) and Rabbit mAb IgG control (ab172730, right panel).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

• FuncS (Neut/Block)

Supplier Data

Functional Studies (Neut/Block) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Cell proliferation of TF-1 (human erythroleukemia erythroblast) cells treated with IL-6 (ab259381) and anti-IL-6 antibody (ab323372) or isotype IgG (ab210849).​

Cell proliferation (%) (A) confirms that the anti-IL-6 antibody (ab323372) neutralizes IL-6-mediated cell proliferation. The IC50 as determined by the dose-dependent inhibition of TF-1 cells proliferation is 4.990 ng/ml (B). ​

TF-1 cells were seeded at 5 × 10⁴ cells per well in a 96-well plate and cultured in medium containing 5% FBS for 48 h. Proliferation was analysed by luminescent assay and recorded using a Microplate Reader at 560nm emission.

• FuncS (Neut/Block)

Supplier Data

Functional Studies (Neut/Block) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Cell proliferation of TF-1 (human erythroleukemia erythroblast) cells treated with IL-6 (ab259381) and anti-IL-6 antibody (ab322419) or isotype IgG (ab210849) Cell proliferation (%) (A) confirms that the anti-IL-6 antibody (ab322419) neutralizes IL-6-mediated cell proliferation. The IC50 as determined by the dose-dependent inhibition of TF-1 cells proliferation is 1.450 μg/ml (B). ​ TF-1 cells were seeded at 5 × 10⁴ cells per well in a 96-well plate and cultured in medium containing 5% FBS for 48 h. Luminescence was recorded using a Microplate Reader at 560nm emission. ​

• FuncS (Neut/Block)

Supplier Data

Functional Studies (Neut/Block) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Cell proliferation of TF-1 (human erythroleukemia erythroblast) cells treated with IL-4 (ab269147) and anti-IL-4 antibody (ab323477) or isotype IgG (ab210849).

Cell proliferation (%) (A) confirms that the anti-IL-4 antibody (ab323477) neutralizes IL-4-mediated cell proliferation. The IC50 as determined by the dose-dependent inhibition of TF-1 cells proliferation is 0.0164 ug/ml (B).

TF-1 cells were seeded at 5 × 10⁴ cells per well in a 96-well plate and cultured in medium containing 5% FBS for 48 h. Proliferation was analysed by luminescent assay and recorded using a Microplate Reader at 560nm emission.

• FuncS (Neut/Block)

Supplier Data

Functional Studies (Neut/Block) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Cell proliferation of TF-1 (human erythroleukemia erythroblast) cells treated with IL-6 (ab259381) and anti-IL-6R antibody (ab323483) or isotype IgG (ab210849).

Cell proliferation (%) (A) confirms that the anti-IL-6R antibody (ab323483) neutralizes IL-6-mediated cell proliferation. The IC50 as determined by the dose-dependent inhibition of TF-1 cells proliferation is 0.1914 ug/ml (B).

TF-1 cells were plated at a density of 5 × 10⁴ cells per well in a 96-well plate. A dilution series of the anti-IL-6R antibody (ab323483) was added to the cells and pre-incubated for 2 hours before IL-6 (ab259381) stimulation, and cells were cultured in medium with 5% FBS for a duration of 48 hours. Proliferation was analysed by luminescent assay and recorded using a Microplate Reader at 560nm emission.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

This data was developed using ab172730 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human spleen labelling Rb IgG with ab172730 at a concentration of 5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins. ab172730 Rabbit IgG, monoclonal [EPR25A] - Isotype Control was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

This data was developed using ab172730 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human liver labelling Rb IgG with ab172730 at a concentration of 5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins. ab172730 Rabbit IgG, monoclonal [EPR25A] - Isotype Control was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

This data was developed using ab172730 the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human heart labelling Rb IgG with ab172730 at a concentration of 5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins. ab172730 Rabbit IgG, monoclonal [EPR25A] - Isotype Control was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IP

Unknown

Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

ab124962 (purified) at 1/20 immunoprecipitating IL-1RA in NIH/3T3 whole cell lysate.
Lane 1 (input) : NIH/3T3 whole cell lysate (10μg)
Lane 2 (+) : ab124962 + NIH/3T3 whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab124962 in NIH/3T3 whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Blocking buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Immunoprecipitation - Anti-IL-1RA antibody [EPR6483] (<a href='/en-us/products/primary-antibodies/il-1ra-antibody-epr6483-ab124962'>ab124962</a>)

All lanes:

NIH/3T3 whole cell lysate

false

• WB

Supplier Data

Western blot - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

Negative Control.

Blocking and dilution buffer and concentration : 5% NFDM/TBST

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 0.5 µg, 1 µg or 2µg of ab172730 [EPR25A]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. H3K4me3 (ab213224) used for comparison. Additional screenshots of mapped reads can be found in the Protocol booklet in the Product Protocol section. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Overlay histogram showing SH-SY5Y (human neuroblastoma) cells stained with ab179513 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab179513 at 1/150 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue with unpurified Rabbit IgG ab172730 at 1/10. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Immunofluorescent staining of HeLa cells using anti-AIF RabMAb (ab32516, left panel) (green) and Rabbit mAb IgG control (ab172730, right panel). DAPI nuclear staining (blue).

Conjugated versions are available for this clone : Alexa Fluor^® 488 (ab199091), Alexa Fluor^® 647 (ab199093), R-PE (ab209478), APC (ab232814).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Immunocytochemistry/immunofluorescence analysis of HeLa cells with purified Rabbit IgG ab172730 at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue with purified Rabbit IgG ab172730 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Overlay histogram showing A549 (human lung carcinoma) cells stained with ab185633 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab185633 at 1/150 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Overlay histogram showing K562 (human chronic myelogenous leukemia) cells stained with ab196018 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab196018 at 1/150 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-Vimentin RabMAb (ab92547, left panel) (brown) and Rabbit mAb IgG control (ab172730, right panel).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

• Flow Cyt

Lab

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Overlay histogram showing A549 (human lung carcinoma) cells stained with ab133557 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab133557 at 1/60 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.

Conjugated versions are available for this clone : Alexa Fluor^® 488 (ab199091), Alexa Fluor^® 647 (ab199093), R-PE (ab209478), APC (ab232814).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

Immunocytochemistry/immunofluorescence analysis of HeLa cells with unpurified Rabbit IgG ab172730 at 1/10. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

• IP

Lab

Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (AB210849)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab172730).

LINE-1 ORF1p was immunoprecipitated from 0.35 mg F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate with ab216324 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216324 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : F9 whole cell lysate 10 μg (Input).
Lane 2 : ab216324 IP in F9 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab216324 in F9 whole cell lysate.

Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : 30 seconds.

false

Exposure time: 30s